Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-kappaB and the expression of chemokine MOB-1, MCP-lgenes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-kappaB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10(-3) mol/L, 10(-4) mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10(-3) mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-kappaB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10(-3) mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10(-5) mol/L atropine) or NF-kappaB inhibitor (10(-2) mol/L PDTC) could obviously inhibit the activation of NF-kappaB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-lgenes in pancreatic acinar cells in vitro through the activation of NF-kappaB.

OBJECTIVETo explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSTotally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).

CONCLUSIONHirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.

Animals ; Aorta ; Apolipoproteins E ; metabolism ; Atherosclerosis ; C-Reactive Protein ; Cholesterol ; Diet, High-Fat ; Drugs, Chinese Herbal ; E-Selectin ; Hirudins ; metabolism ; Interleukin-6 ; Lipids ; Lipoproteins, HDL ; Lipoproteins, LDL ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; metabolism ; Recombinant Proteins ; metabolism ; Triglycerides

OBJECTIVETo observe the clinical effect of chronic fatigue syndrome (CFS) treated with moxibustion at Gaohuang (BL 43).

METHODSWith stratified block randomization, 72 patients accorded with inclusive criteria were divided into a moxibustion at Gaohuang (BL 43) group (moxibustion group) and an acupuncture group, 36 cases in each one. In the moxibustion group, Gaohuang (BL 43) was treated with big moxa cones as the main acupoint, 10 cones a time; Qihai (CV 6) and Zusanli (ST 36) were added with big moxa cones, 7 cones a time. In the acupuncture group, acupoints were the same as those in the moxibustion group, and twirling reinforcing method was used after qi arriving, 60 times one minute and 360° with range. In the two groups, 10-day treatment was made into one course and there were two days between courses. The treatment was given once a day for 3 courses. Changes of fatigue assessment index (FAI) before and after treatment and clinical effects were observed.

RESULTSThe total effective rate was 88.9% (32/36) in the moxibustion group, which was better than 72.2% (26/36) in the acupuncture group apparently (P < 0.05). After treatment in the two groups, FAI scores were obviously declined compared with those before treatment (both P < 0.01) and FAI score in the moxibustion group was apparently lower than that in the acupuncture group (P < 0.05).

CONCLUSIONMoxibustion at Gaohuang (BL 43) can improve the FAI score of patients with CFS and the clinical efficacy is definite.

Acupuncture Points ; Adult ; Fatigue Syndrome, Chronic ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the clinical effect of the sacrococcygeal space injection for the treatment of failed back surgery syndrome.

METHODSFrom July 1998 to October 2012,47 patients with failed back surgery syndrome were treated and included 39 males and 8 females with an average age of 61.5 years old ranging from 35 to 89 years old. Among them,41 patients experienced one time of operation, 6 patients with twice of operation. Forty-one patients underwent single,bilateral fenestration or central laminectomy decompression, discectomy. Six patients underwent total laminectomy discectomy and inter body fusion and pedicle screw fixation. All patients were examined by X-ray plain film, CT or MRI before treatment. The anticoagulation was discontinuation before treatment. The needle was put into the sacrococcygeal gap at prone position in the sense of frustration,suction without cerebrospinal fluid and blood,with injection of Mailuoning (Chinese characters: see text) 15 ml. The pain was assessed by VAS before and after treatment. The Oswestry low back pain disability index and survival quality interference degree were evaluated.

RESULTSAt 1 month after treatment,the pain VAS decreased from 59.24 +/- 17.35 before treatment to 19.19 +/- 11.19 after treatment (P < 0.05); The Oswestry low back pain disability index decreased from (41.35 +/- 9.87)% before treatment to (23.17 +/- 17.56)% after treatment (P < 0.05); The survival quality interference degree decreased from 6.5 +/- 2.2 before treatment to 2.6 +/- 1.4 after treatment (P < 0.05).

CONCLUSIONThe sacrococcygeal gap injection for treatment of failed back surgery syndrome has advantages of simple, safe, fewer complications, and low treatment cost.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Failed Back Surgery Syndrome ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Sacrococcygeal Region ; diagnostic imaging

Objective To compare the clinical outcome of minimally invasive transforaminal lumbar interbody fusion (M-TLIF) and Wiltse-approach TLIF (W-TLIF) in treating single-level degenerative lumbar disease.Methods A retrospective review was performed on the 57 patients with single-level degenerative lumbar disorder managed via M-TLIF (n =27) and W-TLIF (n =30) from December 2009 to December 2010.In M-TLIF group degeneration at the L4-5 disc were noted in 11 cases and at the L5-S1 disc in 16 cases.And 19 cases were diagnosed with lumbar isthmus spondylolisthesis (17 with Grade Ⅰ spondylolisthesis and 2 with Grade Ⅱ spondylolisthesis),4 lumbar spinal stenosis and instability,2 lumbar disc herniation combined with huge posterior osteophytes,1 recurrent lumbar disc herniation after lumbar fenestration,and 1 recurrent lumbar spinal stenosis after decompression.In W-TLIF group degeneration at L4～5 disc were noted in 12 cases and at the L5-S1 disc in 18 cases.There were 19 cases diagnosed with lumbar isthmus spondylolisthesis (18 with Grade Ⅰ spondylolisthesis and 1 with Grade Ⅱ spondylolisthesis),3 with lumbar disc herniation,and 8 with lumbar spinal stenosis.Japanese Orthopedic Association (JOA) score and Visual Analogue Scale (VAS) were used to measure low back and leg pain.Modified Brantigan score was used to assess lumbar interbody fusion.Results Operative time was not significantly different between the two groups (P ＞ 0.05).Incision length and mean blood loss were (5.1 ± 0.7) cm and (90.1 ± 10.5) ml in M-TLIF group,but were (6.9 ± 1.0)cm and (155.3 ±21.2)ml in W-TLIF group (P＜0.05).At postoperative 1 and 3 days VAS in M-TLIF group was (2.1 ± 0.5) points and (1.0 ± 0.1) points respectively,but in W-TLIF group was (3.6 ± 0.1) points and (2.4 ± 1.0) points respectively (P ＜ 0.05).Intraoperative X-ray fluoroscopy frequencies were (46 ± 9) times in M-TLIF group and (7 ± 2) times in W-TLIF group (P ＜ 0.05).Mean period of follow-up was 26.7 months (range,24-36 months).At final follow-up,JOA score,VAS for leg pain and lumbar interbody fusion rate revealed no significant differences between the two groups (P ＞ 0.05),but VAS for lumbar pain was (1.0 ± 0.2) points in M-TLIF group versus (1.9 ± 0.3) points in W-TLIF group (P ＜0.05).Twenty-four cases (89％) had Brantigan score of 3 or over in M-TLIF group and 27 cases (90％) in W-TLIF group,indicating a similar interbody fusion rate (P ＞ 0.05).Conclusions Both lumbar fusion methods are effective in treatment of lumbar degenerative disease.M-TLIF lumbar fusion results in small amount of bleeding,small incision and significantly improved lower back pain as compared with W-TLIF,but W-TLIF involves less exposure to the X-rays.

Objective To evaluate the inhibitive effect of tea polyphenol on the growth of human nasopharyngeal carcinoma HONE1 cell xenograft in nude mice ,and to explore the underlying mechanisms .Methods Tumor model was established by subcu‐taneous inoculation of nasopharyngeal carcinoma cell HONE1 into nude mice ,was used to evaluate the antitumor effect of tea poly‐phenol in vivo .The expression levels of VEGF were detected by real‐time PCR and western blot .Results The growth of xenograft in nude mice was significantly suppressed after application of tea polyphenol at a dose‐dependent manner .To compare with control group ,the inhibition rates were 18 .82% (P<0 .05) and 47 .66% (P<0 .05)when treated at low and high dose respectively ,With in‐creased concentration of TP ,the inhibition rates increased .Real‐time fluorescence quantitative‐PCR and western blot results showed that the expression of VEGF decreased at a dose‐dependent manner .The change of high dose group was obviously ,the difference was statistically significant(P<0 .05) .Conclusion Tea polyphenol could significantly inhibit the growth of human nasopharyngeal carcinoma HONE1 cell xenograft in nude mice ,probably by down regulating the VEGF protein level to inhibit tumor angiogenesis effects .

Objective To evaluate the anticonvulsant effect of midazolam and diazepam when administered nasally on maximal electroshock seizure and metrazol seizure threshold test models .Methods Rats were randomly divided into 7 groups:model group, low-dose, middle-dose and high-dose of midazolam nasal spray groups , diazepam nasal spray in low-dose, middle-dose and high-dose of midazolam nasal spray groups .After the establishment of the maximal electroshock seizure( MES) and metrazol seizure threshold test models ( MST) in rats, the anticonvulsant effects of different doses of midazolam and the clinically used antiepileptic drug diazepam were evaluated and compared .HE staining was used to observe the histopathological changes in the hippocampus , cortex and amygdala in rats .Results Significant anticonvulsant effects were observed on MES and MST in rats pretreated with different dosages of midazolam .In addition , the anticonvul-sant effects of midazolam were stronger than those of diazepam at the same dosage on MES and MST (P<0.05,P<0.01). Histopathological results showed that both midazolam and diazepam could effectively prevent the seizure -induced brain inju-ries, inhibit the increase of microglial cells and the inflammatory cell infiltration in the hippocampus, cortex and amygdala, and reduce the nucleus pycnosis and neuronophagia .Conclusion Midazolam has significantly anticonvulsant and neuropro-tective effects on different seizure models when administered nasally in rats .

Objective To assess the psychometric properties of the Chinese Heartburn Version of Quality of Life in Reflux and Dyspepsia Questionnaire (QOLRAD) in patients with gastroesophageal reflux disease.Methods 130 patients with symptoms of heartburn completed the Chinese version of QOLRAD,the Short-Form-36 (SF-36).30 of them received proton pump inhibitors (PPI) for 8 weeks,which was used to test responsiveness of the Chinese heartburn version of QOLRAD.Results The Chinese version of QOLRAD had acceptable internal consistency.The overall Cronbach's alpha was 0.89 and the internal consistency of dimensions ranged from 0.70～0.90.Content validity index (CVI) was 0.82.Confirmation factor analysis revealed a 5 factor solutions accounting for 62.02％ and most of items in their dimensions had acceptable loads (＞0.4).There was acceptable concurrent validity with correlations between the Chinese heartburn version of QOLRAD and Short Form-36 health survey ranging from 0.172～0.613.As to responsiveness,after therapy of PPI for 8 weeks,except the dimension of sleep disturbance,scores for dimensions of vitality,food/drink problems,physical/social functioning,emotional distress had significant changes.Conclusions The Chinese version of QOLRAD has a good reliability,validity and responsiveness to therapy,which can be used to assess the quality of life in patients with gastroesophageal reflux disease.

Objective To explore Toll like receptor 2 (TLR2) expression after partial liver ischemia/ reperfusion (I/R) in BALB/c mice and its relationship with liver function impairment. Methods All the animals were randomly divided into ischemia/reperfusion injury (I/R) and sham operation (S) groups. Total RNA was extracted from the liver samples and the expression of TLR2 mRNA was analyzed quantitatively by real time PCR method. Membrane protein was extracted, and the expression of TLR2 protein was detected by western blot method. Portal vein serum and plasma were taken for further detection of the levels of alanine aminotransferase (ALT), tumor necrosis factor alpha (TNF ?) and endotoxin . Results After 1h of partial liver ischemia and 4 h of reperfusion, the expression of TLR2 mRNA and TLR2 protein were both remarkably upregulated in tissues of ischemic liver lobes in I/R group compared with that in S group (value of TLR2mRNA: 1.06?0.91vs5.08?1.32,P0.05). Conclusions TLR2mRNA and TLR2 protein expression were upregulated in tissues of ischemic liver lobes after partial I/R injury in mice, and were associated with increased levels of portal vein ALT and TNF ?,and impairment of liver function.

AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.